Print Email Facebook Twitter Determination of denaturated proteins and biotoxins by on-line size-exclusion chromatography-digestion-liquid chromatography-electrospray mass spectrometry Title Determination of denaturated proteins and biotoxins by on-line size-exclusion chromatography-digestion-liquid chromatography-electrospray mass spectrometry Author Carol, J. Gorseling, M.C.J.K. de Jong, C.F. Lingeman, H. Kientz, C.E. van Baar, B.L.M. Irth, H. TNO Defensie en Veiligheid Publication year 2005 Abstract A multidimensional analytical method for the rapid determination and identification of proteins has been developed. The method is based on the size-exclusion fractionation of protein-containing samples, subsequent on-line trypsin digestion and desalination, and reversed-phase high-performance liquid chromatography-electrospray mass spectrometry detection. The present system reduces digestion times to 20 min and the total analysis time to less than 100 min. Using bovine serum albumin and myoglobin as model proteins, optimization of key parameters such as digestion times and interfacing conditions between the different pretreatment steps was performed. The automated system was tested for the identification of infectious disease agents such as cholera toxin and staphylococcal enterotoxin B. This resulted typically in a positive identification by a total sequence coverage of approximately 40%. © 2005 Elsevier Inc. All rights reserved. Subject Biological toxinsLiquid chromatographyMass spectrometryOn-line digestionSize-exclusion chromatographyBovine serum albuminCholera toxinMyoglobinStaphylococcus enterotoxinArticleDesalinationDigestionElectrospray mass spectrometryGel permeation chromatographyNonhumanPriority journalProtein analysisProtein determinationReversed phase high performance liquid chromatographyAmino Acid SequenceCholera ToxinChromatography, GelChromatography, LiquidEnterotoxinsMolecular Sequence DataProtein DenaturationProteinsSpectrometry, Mass, Electrospray IonizationBovinaeChemicals / CAS: Cholera Toxin, 9012-63-9; enterotoxin B, staphylococcal, 39424-53-8; Enterotoxins; Proteins To reference this document use: http://resolver.tudelft.nl/uuid:c538060d-6cf8-4836-9567-b07d4feb259c DOI https://doi.org/10.1016/j.ab.2005.08.023 TNO identifier 238786 ISSN 0003-2697 Source Analytical Biochemistry, 346 (1), 150-157 Document type article Files To receive the publication files, please send an e-mail request to TNO Library.